Assessment of carbon footprint of milk production and identification of its major determinants in smallholder dairy farms in Karnataka, India.

Indian dairy enterprise is dominated by smallholder dairy farms that contribute 72% of the country's total milk production. These smallholder dairy farms are often considered to emit substantial greenhouse gases (GHG) but are poor in productive performances. Therefore, it is crucial to estimate the carbon footprint (CF) of milk production of the smallholder Indian dairy farms. The primary objectives of the study were (1) Assessing the CF of milk production of smallholder dairy farms through life cycle analysis in south-interior Karnataka, India; (2) Identifying the hotspots of GHG emissions and significant factors influencing the CF of milk production in smallholder dairy production system. The study accounted GHG emissions from different sources and considered multiple functions of the smallholder production system. Estimations were made based on primary data collected from 47 farms and associated secondary data. For estimating the CF of milk production, the emissions of carbon dioxide (CO2), methane (CH4), and nitrous oxide (N2O) on a CO2-equivalent (CO2-eq) basis from feed production, enteric fermentation, manure management, transport and energy usage were allocated to fat- and protein-corrected milk (FPCM) based on mass balance, price (crop byproducts and residues) and feed digestibility. Principal component analysis and stepwise linear regression analysis were performed to identify the major factors influencing the CF. The average total GHG emissions (kg CO2-eq yr-1 farm-1) attributable to milk production based on mass, economic, and digestibility allocations were 8,936, 8,641, and 8,759, respectively. The contributions of CH4, N2O, and CO2 to the total farm GHG emission were 70.6%, 20.5%, and 7.69%, respectively. The major emission hotspots were CH4 emission from enteric fermentation (66.8%) and GHG emission from feed production (23.0%). The average CF of cradle-to-dairy cooperative milk production varied from 1.45 to 1.81 kg CO2-eq kg FPCM-1. The CF of milk production was more than 2-fold greater, when milk yield was below 3,500 kg lactating cow-1 yr-1. The FPCM yield 100 kg body weight-1, dry matter intake, and CH4 emission from manure management were the strongest determinants of the CF and explained 83.4% of the observed variation. The study emphasized the importance of considering multiple functions of a mixed crop-livestock-based dairy production system for estimating CF per unit of product. The results suggest that maintaining high-yielding dairy animals and adopting appropriate feeding strategies for better feed utilization are the possible effective interventions for reducing the CF of milk production.

Effect of silkworm (Bombyx mori) pupae oil supplementation on enteric methane emission and methanogens diversity in sheep.

In vitro and in vivo studies were conducted to examine the effect of silkworm pupae oil on methane (CH4) emission and methanogens diversity. Five graded levels (2, 4, 6, 8 and 10%) of silkworm pupae oil were tested in vitro. Eighteen Mandya adult sheep were divided into three groups. All the animals were fed on similar basal diet except the oil supplementation in test groups. Oil level for supplementation was decided on the basis of in vitro study. In vitro study indicated a reduction of 22% in CH4 production with 2% oil supplementation. Animals in test groups were supplemented with oil (2%) either daily (CON) or intermittently (INT) on every alternate week for all the seven days. A significant reduction of 17-20% in enteric CH4 emission (g/d) was achieved due to oil supplementation in sheep. However, No variation was established between test groups CON and INT. In present study, Methanobrevibacter was major genus contributed ∼90% of the total rumen methanogens; whilst Methanobrevibacter gottschalkii was the most abundant methanogens species. Abundance of Methanobrevibacter ruminantium was affected with the oil supplementation. It can be concluded that the silkworm pupae oil at 2% can decrease CH4 emission by 15-20%.

Diversity of rumen microbiota using metagenome sequencing and methane yield in Indian sheep fed on straw and concentrate diet.

An in vivo study aiming to investigate the rumen methanogens community structure was conducted in Mandya sheep fed on straw and concentrate diet. The ruminal fluid samples were collected and processed for unravelling the rumen microbiota and methanogens diversity. Further, the daily enteric methane emission and methane yield was also quantified using the SF6 tracer technique. Results indicated that the Bacteroidetes (∼57%) and Firmicutes (25%) were two prominent affiliates of the bacterial community. Archaea represented about 2.5% of the ruminal microbiota. Methanobacteriales affiliated methanogens were the most prevalent in sheep rumen. The study inveterate that the ruminal archaea community in sheep is composed of 9 genera and 18 species. Methanobrevibacter represented the largest genus of the archaeome, while methylotrophs genera constituted only 13% of the community. Methanobrevibacter gottschalkii was the prominent methanogen, and Methaobrevibacter ruminantium distributed at a lower frequency (∼2.5%). Among Methanomassiliicoccales, Group 12 sp. ISO4-H5 constituted the most considerable fraction (∼11%). KEGG reference pathway for methane metabolism indicated the formation of methane through hydrogenotrophic and methylotrophic pathways, whereas the acetoclastic pathway was not functional in sheep. The enteric methane emission and methane yield was 19.7 g/d and 20.8 g/kg DMI, respectively. Various species of Methanobrevibacter were differently correlated, and the distribution of hydrogenotrophic methanogens mainly explained the variability in methane yield between the individual sheep. It can be inferred from the study that the hydrogenotrophic methanogens dominate the rumen archaeal community in sheep and methylotrophic/aceticlastic methanogens represent a minor fraction of the community. Further studies are warranted for establishing the metabolic association between the prevalent hydrogenotrophs and methylotrophs to identify the key reaction for reducing methane emission.

Comparison of enteric methane yield and diversity of ruminal methanogens in cattle and buffaloes fed on the same diet.

An in vivo study was conducted to compare the enteric methane emissions and diversity of ruminal methanogens in cattle and buffaloes kept in the same environment and fed on the same diet. Six cattle and six buffaloes were fed on a similar diet comprising Napier (Pennisetum purpureum) green grass and concentrate in 70:30. After 90 days of feeding, the daily enteric methane emissions were quantified by using the SF6 technique and ruminal fluid samples from animals were collected for the diversity analysis. The daily enteric methane emissions were significantly greater in cattle as compared to buffaloes; however, methane yields were not different between the two species. Methanogens were ranked at different taxonomic levels against the Rumen and Intestinal Methanogen-Database. The archaeal communities in both host species were dominated by the phylum Euryarchaeota; however, Crenarchaeota represented <1% of the total archaea. Methanogens affiliated with Methanobacteriales were most prominent and their proportion did not differ between the two hosts. Methanomicrobiales and Methanomassillicoccales constituted the second largest group of methanogens in cattle and buffaloes, respectively. Methanocellales (Methanocella arvoryza) were exclusively detected in the buffaloes. At the species level, Methanobrevibacter gottschalkii had the highest abundance (55-57%) in both the host species. The relative abundance of Methanobrevibacter wolinii between the two hosts differed significantly. Methanosarcinales, the acetoclastic methanogens were significantly greater in cattle than the buffaloes. It is concluded that the ruminal methane yield in cattle and buffaloes fed on the same diet did not differ. With the diet used in this study, there was a limited influence (<3.5%) of the host on the structure of the ruminal archaea community at the species level. Therefore, the methane mitigation strategies developed in either of the hosts should be effective in the other. Further studies are warranted to reveal the conjunctive effect of diet and geographical locations with the host on ruminal archaea community composition.

Gastrointestinal nematode and Eimeria spp. infections in dairy cattle along a rural-urban gradient.

Endoparasite infections can lead to considerable economic losses in dairy cattle due to decreases in milk yield and quality. Environmental and host-related factors contribute to endoparasite infection intensity and probability. Moreover, advancing urbanization influences parasite infection dynamics in livestock due to close human-animal cohabitation and changes in animal housing conditions. The aim of the present study was to investigate social-ecological effects on gastrointestinal nematode (GIN) and Eimeria spp. infections in dairy cattle along a rural-urban gradient in the emerging Indian megacity Bangalore. In this regard, 726 faecal samples from 441 dairy cattle of different ages and physiological stages were collected from 101 farms and examined at three visits between June 2017 and April 2018. Based on a survey stratification index (SSI) comprising built-up density and distance to the city center, we assigned the farms to urban, mixed and rural areas. GIN eggs were identified in the faeces of 243 cattle (33.5%; 95% confidence interval [CI]: 30.1-36.9%), and Eimeria spp. oocysts in the faeces of 151 cattle (20.8%; 95% CI: 17.9-23.7%). Co-infection rates of GIN and Eimeria spp. were 8.5 to 12.2% higher in rural compared to urban and mixed areas. The SSI effect significantly influenced Eimeria spp. infection probability and oocyst per gram of faeces (OpG; P < 0.001) with an infection probability and OpG higher than 26% and 40% for cattle kept in rural areas compared to cattle from urban areas. However, the SSI effect was not significant for the infection probability of GIN and for GIN eggs per gram of faeces (EpG). Infection probabilities and EpG/OpG were significantly higher in calves and heifers compared to lactating and dry cows. Moreover, we estimated significantly lower OpG values in summer compared to the other seasons. No differences were estimated for GIN and Eimeria spp. infection probabilities and EpG/OpG with regard to pasture access and breed. The variations in endoparasite infection intensity and probability observed along the rural-urban gradient of Bangalore reflect the variability in dairy husbandry systems governed by the social-ecological context.

Effect of dietary supplementation with phytogenic blend on growth performance, apparent ileal digestibility of nutrients, intestinal morphology, and cecal microflora of broiler chickens.

The effects of dietary supplementation with phytogenic blend (PB) of Aerva lanata, Piper betle, Cynodon dactylon, and Piper nigrum on growth performance, ileal nutrient digestibility, intestinal morphology, and cecal microflora were determined in a 42-day broiler feeding trial. A total of 192 broilers were assigned to 4 dietary treatments (6 replicates and 8 birds/replicate): basal diet, basal diet supplemented with antibiotic (chlortetracycline), 1% and 2% PB, respectively. The body weight gain (BWG) of starter chicks increased linearly (P = 0.023) as dietary supplementation levels of PB increased. At grower phase, broilers fed diet supplemented with 1% PB had similar BWG with the antibiotic group, but other treatments had reduced (P = 0.0001) BWG. Dietary supplementation with 1% PB resulted in the highest (P < 0.0001) BWG during the study. Feed intake was not affected by the treatments during the starter, finisher, and overall rearing periods. Broilers fed diet supplemented with 1% PB had the best (P < 0.0001) feed conversion ratio during the study. Overall, broilers fed only basal diet had the highest (P = 0.0450) mortality. Ileal organic matter (OM) digestibility increased linearly (P = 0.044) with broilers fed diet supplemented with PB, but reduced with antibiotic group. Dietary supplementation with 1% PB had the highest (P = 0.0402) ileal digestibility of tryptophan. In the duodenum, broilers fed diet supplemented with PB had longer (P = 0.0006) villi heights than the birds fed only basal diet, but similar with antibiotic group. Broilers fed diet supplemented with PB had longer (P = 0.0064) villi height in the jejunum than the antibiotic group. Bifidobacterium concentration of the cecum content showed a slight increase (P = 0.053) with increasing supplementation levels of PB. In conclusion, the current study shows that dietary supplementation with PB improves growth performance, intestinal morphology, and apparent ileal digestibility of OM and tryptophan in a dose-dependent manner with the best response at 1% inclusion level.

Characterization of a species-specific repetitive DNA from a highly endangered wild animal, Rhinoceros unicornis, and assessment of genetic polymorphism by microsatellite associated sequence amplification (MASA).

We have cloned and sequenced a 906bp EcoRI repeat DNA fraction from Rhinoceros unicornis genome. The contig pSS(R)2 is AT rich with 340 A (37.53%), 187 C (20.64%), 173 G (19.09%) and 206 T (22.74%). The sequence contains MALT box, NF-E1, Poly-A signal, lariat consensus sequences, TATA box, translational initiation sequences and several stop codons. Translation of the contig showed seven different types of protein motifs, among which, EGF-like domain cysteine pattern signatures and Bowman-Birk serine protease inhibitor family signatures were prominent. The presence of eukaryotic transcriptional elements, protein signatures and analysis of subset sequences in the 5' region from 1 to 165nt indicating coding potential (test code value=0.97) suggest possible regulatory and/or functional role(s) of these sequences in the rhino genome. Translation of the complementary strand from 906 to 706nt and 190 to 2nt showed proteins of more than 7kDa rich in non-polar residues. This suggests that pSS(R)2 is either a part of, or adjacent to, a functional gene. The contig contains mostly non-consecutive simple repeat units from 2 to 17nt with varying frequencies, of which four base motifs were found to be predominant. Zoo-blot hybridization revealed that pSS(R)2 sequences are unique to R. unicornis genome because they do not cross-hybridize, even with the genomic DNA of South African black rhino Diceros bicornis. Southern blot analysis of R. unicornis genomic DNA with pSS(R)2 and other synthetic oligo probes revealed a high level of genetic homogeneity, which was also substantiated by microsatellite associated sequence amplification (MASA). Owing to its uniqueness, the pSS(R)2 probe has a potential application in the area of conservation biology for unequivocal identification of horn or other body tissues of R. unicornis. The evolutionary aspect of this repeat fraction in the context of comparative genome analysis is discussed.

Dye removal from wastewater using activated carbon developed from sawdust: adsorption equilibrium and kinetics.

Mahogany sawdust was used to develop an effective carbon adsorbent. This adsorbent was employed for the removal of direct dyes from spent textile dyeing wastewater. The experimental data were analysed by the Langmuir and Freundlich models of adsorption. Equilibrium data fitted well with the Langmuir model. The rates of adsorption were found to conform to the pseudo-second-order kinetics with good correlation. The equilibrium adsorption capacity of the sawdust carbon was determined with the Langmuir equation as well as the pseudo-second-order rate equation and found to be >300 mg dye per gram of the adsorbent. The most ideal pH for adsorption of direct dyes onto sawdust carbon was found to be 3 and below. The results indicate that the Mahogany sawdust carbon could be employed as a low cost alternative to commercial activated carbon in the removal of dyes from wastewater.